A convenient procedure for the isolation of intact translatable mRNA by potassium iodide gradient centrifugation.

Abstract

High concentrations of KI were found to efficiently protect RNA against degradation by RNases. When a sufficient amount of solid KI was added to cell lysates or subcellular fractions (9 g per 10 ml), the solutions could be stored at room temperature for several days without measurable degradation of mRNA. Ribonucleic acids were selectively sedimented when these KI-containing solutions were centrifuged at 72,000 x g for 24 h. The RNA pellets were found to be readily dissolved in bidistilled water and the redissolved RNA could be immediately submitted to oligo(dT)-cellulose chromatography to isolate the poly(A)-containing RNA. However, extraction with phenol/chloroform was found to be necessary, if total RNA or poly(A)-minus RNA was to be analysed. This procedure was found to be superior to other methods currently in use - especially with respect to the isolation of intact, translatable high-molecular-weight mRNA.

Cite this paper

@article{Mnzner1989ACP, title={A convenient procedure for the isolation of intact translatable mRNA by potassium iodide gradient centrifugation.}, author={P M{\"{u}nzner and Jan Voigt}, journal={Journal of biochemical and biophysical methods}, year={1989}, volume={18 3}, pages={183-93} }